Mechanisms of Podophyllotoxin-Induced Enterotoxicity: A Multi-Omics Integration of Gut Microbiota, Short-Chain Fatty Acids, and Inflammatory Mediators

Podophyllotoxin (PPT), a lignan extracted from the roots and stems of Podophyllum species, exhibits significant enterotoxicity that limits its clinical application. However, its underlying mechanisms remain unclear. This study aimed to explain the mechanisms by which PPT induces enterotoxicity. Changes in body weight, fecal morphology, toxic phenotypes, and histopathological features were evaluated. Hematoxylin-eosin (HE) 3D reconstruction, 16S rRNA sequencing, targeted short-chain fatty acids (SCFAs) analysis, and inflammatory cytokine assays were performed. Our findings demonstrated that PPT induced pathological alterations in rats, including weight loss, diarrhea, and colonic damage. PPT administration significantly reduced beneficial bacteria such as Lactobacillus, while increasing harmful bacteria such as Escherichia-Shigella. Levels of pro-inflammatory cytokines including tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were increased. Additionally, the expression of undecaprenyl-diphosphate synthase (UPPS) and the production of SCFAs were reduced. These results suggest that PPT may alter gut microbial composition, increase invasion of Escherichia-Shigella in intestinal epithelial cells, promote lipopolysaccharide (LPS) production, enhance the release of pro-inflammatory cytokines such as TNF-alpha and IL-6, and inhibit UPPS expression and SCFAs generation, collectively contributing to enterotoxicity. This study provides novel insights into the mechanisms of PPT-induced enterotoxicity, which is important for the prevention and treatment of PPT toxicity.